• Title/Summary/Keyword: RBL 2H3 cell

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Calcium Movement in Carbachol-stimulated Cell-line (Calcium수송기전에 미치는 Carbachol의 영향)

  • Lee, Jong-Hwa
    • The Korean Journal of Pharmacology
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    • v.31 no.3
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    • pp.355-363
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    • 1995
  • It has been well known that the intracellular calcium concentration $([Ca^{2+}]_i)$ in living cell is very sensitive to live or to survive, but the transmembrane system of calcium ion, especially mechanism of calcium ion movement in unexcitable state has been little elucidated. Though many proposed theories for calcium ion transport have been reported, it is still unclear that how could the sustained maintenance in cytosolic calcium level be done in cell. Since one of possible mechanisms of calcium transport may be related to the acetylcholine receptor-linked calcium channel, author performed experiment to elucidate this mechanism of calcium influx related to cholinergic receptor in ml muscarinic receptor-transfected RBL-2H3 cell-line. 1) The effects of carbachol both on calcium ion influx and on the secretion of hexosaminidase were respectively observed in the manner of time-related or concentration-dependent pattern in this model. 2) The effects of several metal cations on calcium transport were shown in carbachol-induced cell-line. 3) Atropine was administered to examine the relationship between cholinergic receptor and calcium ion influx in this model. 4) PMA (Phorbol 12-myristate 13-acetate) or PTx (Pertussis toxin) was respectively administered to examine the secondary mediator which involved pathway of calcium ion movement in carbachol-induced cell-line. The results of this experiments were as follows; 1) Carbachol significantly stimulated both the calcium influx and the secretion of hexosaminidase in the manner of the concentration-dependent pattern. 2) Atropine potently blocked the effects of carbachol in concentration-response manner. 3) Administered metal cations inhibited the calcium influx in carbachol-stimulated this model to the concentration-related pattern. 4) PMA did not inhibit carbachol-induced secretion of hexosaminidase, but blocked the calcium influx in this cell-line. 5) The suppression of carbachol-induced hexosaminidase secretion was shown in PTx-treated cell -line.

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Experimental Study about the Pathway of Inflammatory Allergic Reaction of Cheonmaec-tang (천맥탕(天麥湯)의 알레르기 염증반응 및 기전탐색에 관한 연구)

  • Kim, Yong-Hyeon;Lee, Seung-Yeon;Kim, Won-Il
    • The Journal of Pediatrics of Korean Medicine
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    • v.24 no.1
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    • pp.93-103
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    • 2010
  • Objectives The purpose of this study was to examine the pathway of anti-allergic effects of Cheonmaec-tang (CMT). Methods We examined the cell viability, $\beta$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inbibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation in RBL-2H3 cells pre-treated by CMT of 2.0 mg/ml, 4.0 mg/ml separately. Results We observed that CMT reduced the secretion of $\beta$-hexosaminidase, TNF-$\alpha$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Furthermore, CMT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions These results show that CMT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. These suggest that CMT could be used as an anti-allergic medicine.

Anti-allergic Effects of Socheongyoug-tang on RBL-2H3 Mast Cell and Mice-mediated Allergy Model (RBL-2H3 비만세포와 동물병태모댈에서 소청룡탕(小靑龍湯)의 항알레르기 효과)

  • Kim, Kyung-Yeol;Lee, Jong-Hwa;Kim, Young-Jun;Choi, Sung-Youl;Kim, Tae-Heon;Lyu, Yeoung-Su;Kang, Hyung-Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1260-1270
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    • 2007
  • The discovery of drugs on the treatment of mast cell-mediated allergic disease is a very important subject in human heath. The Socheongyoug-tang(SCYT) has been used for centruries as a traditional medicine in Korea and is known to have an anti-inflammatory effect. However, its specific mechanism of action is still unknown. In this report, we investigated the effect of hot water extract from SCYT on RBL-2H3 mast cell-mediated allergic reaction and studied its possible mechanism of action. SCYT inhibited compound 48/80-induced systemic anaphylaxis and serum histamine release in mice. SCYT decreased the passive cutaneous anaphylaxis reaction activated by Anti-lgE antibody-HSA. SCYT dose-dependently reduced histamine release from mice peritoneal mast cells activated by Anti-lgE antibody-HSA. SCYT increased cAMP and decreased compound 48/80-induced intracellular $Ca^{2+}$ levels. Our findings provide evidence that SCYT inhibits mast-derived allergic reactions, and also demonstrate the involvement of cAMP and intracellular $Ca^{2+}$ in these effects.

Anti-allergic Effect of Seungmagalgeun-tang through Suppression of NF-${\kappa}B$ and p38 Mitogen-Activated Protein Kinase Activation in the RBL-2H3 Cells

  • Lyu, Ji-Hyo;Lyu, Sun-Ae;Yoon, Hwa-Jung;Ko, Woo-Shin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1572-1578
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    • 2008
  • In previous report, Seungmagalgeun-tang (SGT) could exert its anti-inflammatory actions in the BV-2 microglial cells. However, study on the anti-inflammatory effect of SGT in mast cells has not been identified. Therefore, we examined on the anti-inflammatory effect of SGT on the phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187-induced rat basophilic leukemia (RBL-2H3) cells. SGT inhibited the release of ${\beta}$-hexosaminidase and secretion and expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-4 on RBL-2H3 cells, without affecting cell viability. The protein expression level of nuclear factor (NF)-${\kappa}B$ (p65) was decreased in the nucleus by SGT. In addition, SGT suppressed the degradation of inhibitory protein $I{\kappa}B-{\alpha}$ protein, the activation of p38 mitogen-activated protein kinase (MAPK), and the expressions of cyclooxygenase (COX)-2 mRNA and protein level in RBL-2H3 cells. These results suggest that SGT could be involved anti-allergic effect by control of NF-${\kappa}B$ (p65) translocation into the nucleus through inhibition of $I{\kappa}B-{\alpha}$ degradation and suppression of COX-2 expression.

Anti-allergic Effects of Petasites japonicum (머위(Petasites japonicum) 추출물의 항알레르기 효과)

  • 최옥범
    • The Korean Journal of Food And Nutrition
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    • v.15 no.4
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    • pp.382-385
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    • 2002
  • It is well known that the Petasites japonicum have been used for a long time in traditional medicine for the treatment of allergic diseases such as lacquer poisoning and asthma. Anti-allergic actions of Petasites japonicum extracts were asessed by testing their effects on the degranulation of mast cells. For this, hexosaminidase released (degranulation marker) from RBL-2H3 cells(mast cell line) was used. At the concentration of 300 $\mu\textrm{g}$/mL of the methanol, ethylacetate and hot water extract, the degranulation of RBL-2H3 cells were inhibited 83.33, 69.75 and 35.4%, respectively. These results suggest that the Petasites japonicum could be provide a effective resource for the control of allergic diseases.

Inhibitory Effect of Rhododendron Mucronulatum Root Extract on Allergic Inflammation (진달래 뿌리 추출물의 알레르기 염증 억제 효과)

  • Jang, Si Sung;Lee, DaeJoong;Song, Jihoon;Park, Do Hwi;Jeon, Chan Yong;Hwang, Gwi Seo
    • The Journal of Internal Korean Medicine
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    • v.43 no.1
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    • pp.68-78
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    • 2022
  • Objective: In this study, we investigated the protective effect of rhododendron mucronulatum extract (RME) on allergic reactions and inflammation. Methods: The effect of RME was determined using ELISA and RT-PCR in RBL-2H3 mast cells and RAW 264.7 macrophage cells. We determined cell viability, β-hexosaminidase release, and the synthesis of IL-4 and TNF-α in RBL-2H3 cells. In addition, we determined NO from RAW 264.7 and the gene expression of IL-1β, iNOS, IL-6, TNF-α, and IL-10. Results: RME inhibited β-hexosaminidase release and synthesis of IL-4 and TNF-α in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. In addition, RME inhibited the production of NO and the gene expression of IL-1β, iNOS, IL-6, and TNF-α in LPS-stimulated RAW 264.7 cells. Conclusion: From these results, we concluded that RME possesses anti-allergic activity and anti-inflammatory activity due to the inhibition of mast cells and macrophage function.

Effects of Eucommia ulmoides Oliver Extract on Inhibition of β-hexosaminidase and Keratinocyte Differentiation (β-hexosaminidase 분비 억제 및 각질형성세포 분화에 대한 두충(Eucommia ulmoides Oliver) 추출물의 효과)

  • Hong, In Kee;Kim, Eun Ji;Seok, Ji Hyun;Kim, Bo Hyeon;Jang, Jin Dong;Joe, Gi Jung;Choi, Shin Wook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.1
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    • pp.21-28
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    • 2014
  • In this study, Eucommia ulmoides Oliver extracts was studied in order to see any effects on the ${\beta}$-hexosaminidase release suppression of RBL-2H3 cells and on the expression of filaggrin, transglutaminase-1 (TGase-1) and cornified cell envelope (CE) related to the recovery of HaCaT keratinocyte skin barrier. Results showed that Eucommia ulmoides Oliver extracts reduced ${\beta}$-hexosaminidase release in RBL-2H3 cells and increased the effects of Eucommia ulmoides Oliver extract on the expression of filaggrin, transglutaminase-1 (TGase-1) and cornified cell envelope (CE) in HaCaT keratinocytes. Taken together, these results suggested that Eucommia ulmoides Oliver extract may be applicable for keratinocyte differentiation.

PKC Isotype that Affects the Interaction of HRF with Na, K-ATPase (Na,K-ATPase와 IgE-Dependent Histamine Releasing Factor의 결합에 영향을 미치는 Protein Kinase C Isotype에 관한 연구)

  • Sohn Wern-Joo;Lee Kyunglim
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.260-266
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    • 2005
  • IgE-dependent histamine releasing factor (HRF), previously known as P23/P21 or translationally controlled tumor protein (TCTP), induces the degranulation of histamine in mast cell and basophil. Yeast two hybrid results showed that HRF interacts with the alpha subunit of Na, K-ATPase, suggesting that HRF is a regulator for governing the activity of Na, K-ATPase. In this study, we examined the interaction of HRF and Wa,K-ATPase after treatments of various PKC isotype inhibitors. Membrane fractionation, pull-down assay and immunoprecipitation results showed that PKC $\alpha,\;PKC\;\beta,\;\delta$ subunits are involved in the phosphorylation of HRF. However, these results did not correlate with the results of histamine release assay since histamine release assay results suggested that some PKC isotype inhibitors induced the histamine release in RBL-2H3 cell.

Study on Anti-Allergic Effecst of Ganoderma lucidum Herbal Acupuncture and Ganoderma lucidum Extract (영지(靈芝) 약침(藥鍼)과 영지(靈芝) 추출액의 항알레르기 효과에 대한 연구)

  • Kang, Kyung-Hwa;Youn, Hyoun-Min
    • Journal of Pharmacopuncture
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    • v.10 no.3
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    • pp.37-46
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    • 2007
  • Objectives We studied on anti-allergic effects of Ganoderma lucidum herbal acupuncture(GHA) and Ganoderma lucidum extract(GE). Methods In vivo, Animals were herbal-acupunctured GHA at both B13s three times for 5 days. Then, we investigated compound 48/80-induced active systemic anaphylatic shock using ICR mice and anti-DNP IgE-induced passive cutaneous anaphylaxis using Sprague Dawley rat. In vitro, we measured cell viability, b-hexosaminidase release, IL-4 and TNF-a from RBL-2H3 cells, and nitric oxide from Raw264.7 cell after treatment of GE of various concentrations. Results In vivo, GHA pretreatments at both B13s inhibited compound 48/80-induced active systemic anaphylatic shock. Passive cutaneous anaphylaxis were inhibited by GHA10 and OP. In vitro, $0.1\;{\sim}\;2%$ GE treatments were not affect on cell viability and inhibited b-hexosaminidase release, IL-4, TNF-a and nitric oxide. Conclusions These results suggest that GHA and GE may be beneficial in the inhibition of allergic inflammatory response.

Inhibitory Effect of Dendrobium moniliforme on Degranulation and Histidine Decarboxylase Expression in RBL-2H3 Cells (RBL-2H3 세포에서 탈과립과 histidine decarboxylase 발현에 미치는 석곡(Dendrobium monilifrme)의 효과)

  • Young Ji Lee;Iskander Madhi;YoungHee Kim
    • Journal of Life Science
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    • v.33 no.2
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    • pp.176-182
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    • 2023
  • The stems of Dendrobium moniliforme are used in traditional Oriental medicine as a Yin tonic to nourish the stomach, promote the production of body fluid, and reduce fever. This study investigated the effects of the aqueous extract of D. moniliforme stems (DME) on mast cell degranulation and the expression of tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and histamine-synthesizing enzyme histidine decarboxylase (HDC). We used rat mast cell line RBL-2H3 cells and stimulated them with PMA plus calcium ionophore (PMACI). Pretreatment with DME significantly inhibited PMACI-induced β-hexosaminidase release and the expression of TNF-α, IL-4, and HDC. Furthermore, DME suppressed PMACI-induced nuclear translocation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein 1 (AP-1). In addition, HDC expression was inhibited by SP600125 (JNK inhibitor), PD98059 (ERK inhibitor), and SB203580 (p38 kinase inhibitor). Finally, the phosphorylation of p38 kinase, extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase (JNK) was inhibited by pretreatment with DME. These results suggest that DME has inhibitory effects against degranulation, cytokine (TNF-α and IL-4) and HDC expression, and that HDC expression is mediated by MAPK signaling. These findings suggest that DME may have therapeutic potential in the treatment of hypersensitive and inflammatory diseases.